Gallbladder metastasis from hepatocellular carcinoma: Report of a case and review of literature

INTRODUCTIONThe incidence of metastasis of hepatocellular carcinoma (HCC) to the gallbladder is low. Here, we report a case of HCC with metastasis to the gallbladder and discuss the pattern of spread and the treatment.PRESENTATION OF CASEA 74-year-old man was diagnosed with advanced hepatocellular carcinoma. Computed tomography and magnetic resonance imaging demonstrated a tumor in the right lobe of the liver with a thrombus in the bifurcation of the portal vein. Because intraoperative ultrasonography showed portal vein tumor thrombosis from the main tumor reaching the umbilical portion, we performed only a cholecystectomy for the elimination of postoperative cholecystitis. Pathological examination showed gallbladder vein tumor thrombosis from poorly differentiated hepatocellular carcinoma.DISCUSSIONA preoperative diagnosis of metastatic HCC to the gallbladder is difficult because there are no specific findings in the imaging tests. Cancer cells in the liver were thought to migrate to the gallbladder via the connection between the portal system and the cholecystic veins, and grow in the lumen of the veins in our case. The survival rate, in all reported cases including the present case, was increased in patients who underwent radical resection, compared to patients who underwent palliative surgery.CONCLUSIONThe resection of metastatic HCC to the gallbladder might appear to prolong survival.     

TACE联合RFA治疗原发性肝细胞癌预后多因素分析

目的探讨TACE联合RFA治疗原发性肝癌（HCC）预后的主要影响因素。方法回顾性分析86例经治疗的HCC患者的临床和随访资料,观察远期疗效,应用Kaplan-Meier法计算累积生存率,绘制生存曲线,对可能影响预后的16项因素行Cox多因素回归分析。结果本组患者6、12、24、36个月生存率分别为94.20%、89.40%、65.80%、49.70%,平均生存时间为（42.98±3.80）个月。多因素分析显示影响预后因素包括肝功能Child分级、Barcelona分期、肿瘤最大直径及治疗效果。结论 TACE联合RFA是治疗HCC的有效方法,其中患者肝功能Child分级、肿瘤Barcelona分期、肿瘤大小是影响预后的危险因素,治疗效果是影响预后的保护性因素。     

经肝内注射肿瘤细胞建立小鼠肝癌实验模型的效果观察

目的 探讨经肝内直接注射肝癌细胞建立小鼠肝癌原位模型的效果。方法 选取Balb/c小鼠来源的ML-3肝癌细胞经肝内直接注射接种至肝脏实质内,与经脾注射肝癌细胞相比较,对其肝内成瘤情况进行观察。结果 肿瘤细胞接种第6天起,经脾脏注射组以及经肝脏注射组小鼠均不同程度出现体重减轻、腹水、黄疸以及腹腔可触及包块。在经脾脏注射组,15只小鼠中仅有3只长出肝脏肿块,其余均腹腔种植转移。在经肝脏直接注射组,15只小鼠中有10只长出肝癌,其中伴腹腔种植转移1只。结论 经肝脏直接注射肿瘤细胞较经脾脏注射法能更有效诱导建立肝癌原位模型。     

裸鼠人肝癌原位移植多药耐药模型的建立及其耐药机制的探讨

目的　在体内建立人肝癌裸小鼠原位移植多药耐药模型,并研究其耐药机理。方法采用人肝癌（BEL-7402）裸小鼠（4～6周龄）原位移植,给予阿霉素(1.5mg/kg体重,每周1次)腹腔注射诱导耐药（共8周）,经四唑蓝(MTT)快速比色法检测原代培养的耐药细胞对抗癌药的敏感性,以流式细胞仪检测癌细胞表面mdr1基因产物P-糖蛋白（P-gp）的表达,并用罗丹明试验观察该蛋白功能。结果　移植瘤组织形态及生物学方面符合人肝癌特征,实验组肝癌细胞表面P-gp表达为(75.45±5.67)%,而对照组表达仅(4.25±1.28)%,差异有非常显著性（P＜0.01）,对阿霉素的耐药倍数提高了16.67倍,对羟基喜树碱具有交叉耐受性（13.67倍）。耐药细胞表面P-gp有较强的药物外排功能。结论　阿霉素较易诱导原位移植于裸小鼠的人肝癌多药耐药性的产生,其耐药倍数与临床肝细胞癌相似。该模型的建立对研究肝癌多药耐药的产生机制以及逆转其多药耐药性具有重要价值。     

人肝癌裸鼠NKG2D受体的表达及对NK细胞毒活性的影响

目的 通过研究原发性肝癌中NK细胞的细胞毒活性变化及其活化性受体NKG2D的表达,观察肝脏和脾脏组织病理变化,探讨NK细胞免疫抑制机制,为原发性肝癌的免疫治疗提供理论依据。方法 ①建立人肝癌裸鼠皮下-肝原位移植瘤模型：先用人肝癌细胞株Hep3B接种于裸鼠皮下,形成皮下移植瘤,然后用此移植瘤组织再接种于裸鼠肝内,建立肝原位移植瘤模型（间接肝原位移植瘤模型）;②检测裸鼠原发性肝癌对NK细胞免疫活性的影响：分离裸鼠外周血、肝脏及脾脏组织NK细胞,LDH方法检测NK细胞的细胞毒活性,流式细胞技术检测不同组织NK细胞NKG2D表达百分率,H-E染色观察肝脏移植瘤对肝脏和脾脏淋巴细胞的影响。结果 ①外周血、肝脏及脾脏的NK细胞毒活性及NKG2D受体表达随着肿瘤生长逐渐下降,其中肝脏NK细胞毒活性及NKG2D表达下降明显;②荷瘤裸鼠肝脏癌组织与皮下瘤相同,癌组织异型性与时间呈正相关,脾脏淋巴小结在第4周增生明显,第8周小梁结构增多。结论 原发性肝癌通过下调NKG2D的表达,对NK细胞有免疫抑制作用,这种作用主要发生在肝脏,但对外周血和脾脏也有影响。     

雷帕霉素对人肝癌裸鼠原位移植瘤生长的影响

目的：探讨免疫抑制剂雷帕霉素对肝癌细胞生物学行为的影响。方法：建立肝癌裸鼠原位移植瘤模型80只,随机分为空白对照组、环孢霉素(CsA)处理组、雷帕霉素常规剂量处理组和高剂量处理组,每组20只。用药2周后观察肿瘤生长情况,RT-PCR方法检测肿瘤组织中CD44v6 mRNA的表达。结果：雷帕霉素常规剂量组及高剂量组肿瘤体积较空白对照组明显缩小, CD44v6的表达显著下调(P<0.05);CsA组肿瘤体积较之空白对照组明显增大, CD44v6的表达显著上调(P<0.05)。结论：CsA可促进肝癌的生长和转移;雷帕霉素具有显著抑制肝癌侵袭转移的作用,能抑制肝癌细胞CD44v6基因的表达。     

重组人p53腺病毒注射液瘤内注射联合125I粒子植入治疗肝细胞癌残余灶

目的评价重组人p53腺病毒（rAd-p53）注射液瘤内注射联合125I粒子组织间植入对肝细胞癌（HCC）残余灶的治疗效果。方法 38例经过TACE及物理消融治疗后HCC患者,共发现肝癌残余灶69个。对17例30个病灶进行rAd-p53瘤内注射联合125I粒子植入治疗（联合治疗组）;对21例39个病灶进行单纯125I粒子植入术（单纯125I粒子植入组）。对比观察两组患者肿瘤大小变化及不良反应。结果治疗后1、2和6个月,两组的有效率差异有统计学意义（P〈0.05）;两组间比较,完全缓解、部分缓解、稳定和进展差异无统计学意义。结论 rAd-p53瘤内注射联合125I粒子植入对HCC残余灶的短期治疗效果优于单纯125I粒子植入术。     

四氯化碳/乙醇诱导129/Sv小鼠产生原发性肝细胞癌模型的建立

目的　建立稳定可靠的肝癌动物模型 ,为深入进行肝细胞癌的实验研究打下良好的基础。方法　取 90只 12 9/Sv小鼠 ,采用四氯化碳 /乙醇诱导 2 0周 ,观察其肝癌发生的情况。另取 10只作为正常对照组。结果　 90只 12 9/Sv小鼠共死亡 4 7只 ,最后剩余的 4 3只小鼠中有 16只发生肝癌。而常对照组 10只小鼠 ,其肝脏均未发现明显异常。诱导组小鼠的肝细胞癌类型以中、高分化为主。结论　采用四氯化碳 /乙醇诱发的 12 9/Sv小鼠肝癌 ,模型稳定 ,癌变率高 ,经历了肝炎、肝硬化阶段 ,与人肝癌的发生发展过程非常相似。因此 ,是较理想的研究肝癌的实验动物模型。     

c-kit在人肝硬化及肝细胞肝癌组织中的表达及意义

目的探讨肝脏祖细胞标志物c-kit在人肝硬化及肝细胞肝癌（HCC）组织中的表达及其与临床病理特征的关系。方法对30例肝硬化、40例肝细胞肝癌标本及3例正常组织标本进行常规组织学观察以及c-kit、CD45免疫组化染色,对肿瘤细胞的分化程度及肝硬化组织门静脉炎症程度进行分型和评分,分析c-kit表达与肝硬化及肝细胞肝癌的临床病理特征的联系。结果正常肝脏中c-kit染色阴性。20／30例肝硬化中发现c-kit（＋）细胞,位于门脉周围区域和纤维间隔内,个别阳性细胞整合到成熟胆管,肝硬化结节中没有发现c-kit（＋）细胞。19／40例HCC组织中存在c-kit （＋）肿瘤细胞,在肿瘤细胞之间或肿瘤结节周围分散分布。HBsAg及Anti-HBc在c-kit（＋）与c-kit （-）HCC之间表达有显著性差异（χ^2=5．063,P〈0．05;χ^2=6．667,P〈0．05）。c-kit表达与肿瘤的分化程度紧密相关（χ^2=10．384,P〈0．05）,分化程度越低,c-kit表达越高。结论骨髓来源的肝脏祖细胞参与了部分肝硬化病变过程中的肝再生及HCC的形成和发展,c-kit表达情况对于判断HCC预后有一定意义。     

PTK787/ZK222584 Combined with Interferon Alpha and 5-Fluorouracil Synergistically Inhibits VEGF Signaling Pathway in Hepatocellular Carcinoma

Background

The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus remains poor. We previously reported the beneficial effects of interferon alpha (IFN) and 5-fluorouracil (5-FU) combination therapy for these patients. We showed that the mechanism of therapy was regulation of vascular endothelial growth factor (VEGF). Here, we combined IFN/5-FU therapy with the VEGF receptor–selective inhibitor PTK787/ZK222584 (PTK/ZK) and examined the antitumor effects and the mechanism of action.

Methods

We studied two HCC cell lines, PLC/PRF/5 and HuH7, and a human umbilical vein endothelial cell line, HUVEC. We studied the effects of IFN/5-FU with or without PTK/ZK in growth inhibition assays, immunohistochemistry, Western blot analysis, and immunocytochemistry.

Results

In a HuH7 xenograft model, the combination of PTK/ZK and IFN/5-FU significantly inhibited proliferation, induced apoptosis, decreased microvessel density, reduced the number of tumor cells that expressed VEGF receptor 2 (VEGFR-2), and repressed the phosphorylation of Akt in vivo. In HCC cells and HUVECs in vitro, IFN/5-FU plus PTK/ZK repressed the expression of VEGFR-2 and repressed the phosphorylation of VEGFR, Akt, Erk, and p38MAPK.

Conclusions

VEGF signaling inhibition enhanced the antitumor effects of IFN/5-FU therapy on HCC cells and endothelial cells via Erk, Akt, and p38MAPK pathways.

     

Solitary metastasis to the intercostal muscle from hepatocellular carcinoma: A case report

INTRODUCTIONSkeletal muscle metastases from carcinomas, especially to intercostal muscles, are rare. Most metastatic chest wall tumors from hepatocellular carcinoma (HCC) result from disseminations through needle tracts of intrahepatic HCC treatments.PRESENTATION OF CASEWe report the case of a 65-year-old man with chronic viral hepatitis B whose intrahepatic lesions were stabilized by repeated radiofrequency ablations and transcatheter arterial chemoembolization. Follow-up computed tomography demonstrated a well-enhanced mass in the right chest wall. Because α-fetoprotein and des-γ-carboxy prothrombin levels were elevated and no other tumors were detected, we diagnosed the mass as an extrahepatic metastasis from the HCC and resected it along with the surrounding ribs. There was no involvement of the bone, pleura, and lung.DISCUSSIONThe tumor was microscopically diagnosed as an intercostal muscle tumor metastasized from HCC, which has not been documented previously. The resection rate of extrahepatic tumors of HCC is low in literature. No other apparent extrahepatic recurrence has been observed for more than 20 months after the surgery.CONCLUSIONWe report the case of HCC patient who underwent surgical resection of an intercostal muscle tumor that had metastasized from HCC. Pathological examination of the tumor revealed the tumor cells in the blood vessels, and we speculate it hematogeneous metastasis.     

天花粉蛋白对荷人肝癌裸小鼠的抑癌作用

目的 探讨天花粉蛋白(trichosanthin,TCS)对荷人肝癌裸小鼠的抑癌作用。方法 采用人肝癌原位移植模型。将24只裸小鼠分为TCSⅠ组、TCSⅡ组、TCSⅢ组和对照组，TCS剂量分别为0．2mg／kg、0．4mg／kg、0．6mg／kg，对照组用生理盐水0．2ml／只腹腔注射，共10次。第21天称瘤重和裸小鼠体重，计算抑癌率。应用免疫组织化学技术，检测瘤组织中Ki67的表达。结果 对照组、TCSⅠ组、TCSⅡ组和TCSⅢ组平均瘤重分别为0．99g、0．81g、0．22g和0．20g。用药组瘤重与对照组相比有显著性差异(P＜0．05)。TCSⅠ组、TCSⅡ组和TCSⅢ组抑癌率分别为18．18％、77．78％、79．80％。对照组、TCSⅠ组、TCSⅡ组和TCSⅢ组平均鼠重增加为2．22g、1．22g、0．28g、一2．37g。用药组与对照组鼠重变化有显著性差异(P＜0．05)。对照组、TCSⅠ组、TCSⅡ组和TCSⅢ组Ki67蛋白阳性表达百分率分别为31．33％、29．92％、10．25％和9．67％。TCSⅠ组Ki67百分率与对照组相比无显著性差异(P＞0．05)。TCSⅡ组和TCSⅢ组Ki67百分率与对照组相比有显著性差异(P＜0．05)。结论 TCS对荷人肝癌裸小鼠的移植瘤有明显的抑癌作用。TCS抑制肿瘤细胞增殖可能是其抑癌的机制之一。     

肝癌特异性细胞毒T淋巴细胞的实施及临床研究

诱导肿瘤特异性细胞毒性Ｔ细胞，观察其体外、体内的抗肿瘤作用及其特异性；评价其扩上癌术后复发的临床应用价值。方法用细胞因子体外周期刺激方法提高肝癌细胞的免疫原性，再与肿瘤浸润Ｔ细胞共同培养，辅以ＣＤ２８产市民人刺激，诱导ＴＳ－ＣＴＬｓ，检测其体内外抗肝癌作用及其效应机制；大量扩增后回输患者，观察其免疫学指标的变化随访其术后复发情况。结果，体外研究表明，肝癌细胞经ＩＮＦ－γ、ＴＮＦ－α诱导后ＭＨＣＯＤ     

肝癌姑息性切除术后裸鼠残癌模型的建立

目的 建立肝癌姑息性切除术(PR)后可研究残癌生物学特性变化的转移性裸鼠人肝癌原位移植瘤模型.方法 采用单肝叶双瘤源原位接种技术建立MHCC97H原位移植瘤模型,PR切除1瘤,观察5周.测量肿瘤大小,流式细胞术计数外周血循环肿瘤细胞数,荧光显微镜和苏木素-0伊红(HE)染色检测肝内、肺、腹腔转移等,免疫组织化学染色和Western blot检测脑和肿瘤组织Vimentin表达.结果 双瘤源接种成瘤率95.6％、PR成功率95.3％、裸鼠5周总体存活率90.2％.假手术组、肝部分切除术组、PR组肿瘤大小、循环肿瘤细胞数均依次升高,肝内转移率分别为15.4％、41.7％、83.3％,肺转移率分别为61.5％、91.7％、100.0％,腹腔转移率分别为23.1％、50.0％、83.3％(P值均＜0.05).手术组脑和肿瘤组织Vimentin表达均显著上调.结论 成功建立转移性裸鼠人肝癌原位移植瘤合并PR模型,可用于研究残癌生物学特性的变化.     

肝动脉化疗栓塞术治疗原发性肝细胞癌预后影响因素分析

目的分析肝动脉化疗栓塞术(TACE)治疗原发性肝细胞癌(HCC)的预后影响因素。方法回顾性分析326例接受TACE治疗的HCC患者的资料。采用Kaplan-Meier法计算患者1、2、3年累积生存率,分别以Log-rank检验及Cox比例风险模型进行预后相关单因素及多因素分析。结果患者1、2、3年累积生存率分别为73.90%、40.20%、22.20%,中位生存期21个月。单因素分析显示,甲胎蛋白(AFP)、γ-谷氨酰转肽酶(GGT)、肿瘤最大径、肿瘤数目、肝功能Child-Pugh分级、巴塞罗那临床肝癌(BCLC)分期、门静脉癌栓、动静脉瘘、远处转移为影响HCC患者预后的相关因素(P均0.05)。多因素分析显示,AFP、GGT、肿瘤最大径、肿瘤数目、BCLC分期、动静脉瘘为HCC患者预后的独立影响因素(P均0.05)。结论影响TACE治疗HCC患者预后的独立因素包括AFP、GGT、肿瘤最大径、肿瘤数目、BCLC分期及动静脉瘘。     

KDR启动子驱动的CD/TK双自杀基因联合survivin基因干扰抑制肝癌细胞生长的体内研究

目的：探讨腺病毒介导的KDR启动子驱动的CD/TK双自杀基因系统（Ad-KDRP-CD/TK）联合survivin基因干扰对肝癌细胞在裸鼠体内生长的抑制作用。
　　方法：将20只裸鼠随机分均为模型组（皮下植入BEL-7402肝癌细胞成瘤,不加任何处理）、双自杀基因转染组（皮下植入转染Ad-KDRP-CD/TK的BEL-7402细胞,成瘤后瘤内注射前药更昔洛韦与5-氟胞嘧啶）、survivinsiRNA转染组（皮下注射BEL-7402肝癌细胞,成瘤后瘤内注射survivinsiRNA/Lip-DMEM转染混合物）、联合转染组（双自杀基因转染+survivinsiRNA转染处理）。治疗2周后处死各组小鼠,称取肿瘤质量,计算肿瘤抑制率,检测瘤组织微血管密度（MVD）及survivinmRNA与蛋白表达。结果：各治疗组的肿瘤质量明显小于模型组（均P<0.05）,且联合转染组的抑瘤率最大（均P<0.05）;肿瘤组织MVD、survivinmRNA与蛋白水平均明显降低,且联合转染组的降低程度最为明显（均P<0.05）。
　　结论：双自杀基因联合survivin干扰是抑制鼠肝癌细胞在体内生长的有效途径。     

肝纤维化对射频消融治疗单发肝细胞癌患者长期预后的影响

目的观察肝纤维化程度对射频消融治疗后单发肝细胞癌(HCC)患者长期预后的影响。方法回顾性分析363例美国国立癌症研究所监测、流行病学和结果数据库(SEER)中经射频消融治疗单发HCC患者,根据肝纤维化评分(FS)将其分为0~4分组(FS 0~4分组)和5~6分组(FS 5~6分组)。采用Kaplan-Meier法构建生存曲线,以COX比例风险模型分析影响预后的因素。结果与FS 0~4分组比较,FS 5~6分组患者中位病因特异性生存期(CSS)下降(33个月vs 22个月,P<0.001)。FS 5~6分(P<0.001)、伴血管侵犯(P=0.041)、AFP升高(P=0.042)及肿瘤长径>3 cm(P=0.026)是CSS下降的独立危险因素。结论肝纤维化是影响射频消融治疗后单发HCC患者长期预后的独立危险因素。     

High expression of ADAM8 correlates with poor prognosis in hepatocellular carcinoma

ObjectivesTo evaluate the association between ADAM8 tissue expression and patient prognosis in hepatocellular carcinoma (HCC).MethodsADAM8 expression was analyzed using immunohistochemical staining methods on tissue samples from a consecutive series of 105 HCC patients who underwent resections between 2000 and 2006. The correlation of ADAM8 expression and patients' clinicopathological parameters was evaluated. Survival analysis was performed using the Kaplan–Meier method and Cox's proportional hazards model.ResultsADAM8 was highly expressed in 54.3% of the HCC patients. The ADAM8 expression level was closely associated with serum AFP elevation, tumor size, histological differentiation, tumor recurrence, tumor metastasis, and tumor stage. Kaplan–Meier survival analysis showed that a high expression level of ADAM8 resulted in a significantly poor prognosis of HCC patients. Multivariate analysis revealed that ADAM8 expression level was an independent prognostic parameter for the overall survival rate of HCC patients.ConclusionsThese findings provide evidence that a high expression level of ADAM8 serves as a biomarker for poor prognosis for HCC. Thus, we speculate that ADAM8 may be a potential target of antiangiogenic therapy for HCC.     

Corruption of neuroblastoma patient derived xenografts with human T cell lymphoma

BackgroundPatient derived xenografts (PDXs) provide a unique opportunity for investigators to study tumor cell activity, response to therapeutics, and resistance patterns without exposing the human patient to experimental compounds, and thereby play a crucial role in pre-clinical evaluation of new therapies. It has been reported that PDXs may undergo a transformation to lymphoma, most commonly associated with Epstein Barr virus (EBV). If the character of a xenograft becomes compromised and remains undetected, it could have a detrimental impact on the research community as a whole. Our lab has established a number of pediatric solid tumor PDXs which accurately recapitulate the human tumors following several passages. One particular neuroblastoma PDX was noted to grow quickly and with an unusual phenotype, leading us to hypothesize that this PDX had undergone a transformation.MethodsThe PDX in question was investigated with histology, immunohistochemistry (IHC), EBER in situ hybridization, and PCR to determine its identity.ResultsHistology on the tumor revealed a small, round blue cell tumor similar to the original neuroblastoma from which it was derived. IHC staining showed that the tumor was composed of lymphocytes that were CD3 positive, < 5% CD4 positive, and CD20 negative. The cells were Epstein Barr virus negative. PCR demonstrated that the tumor was human and not murine in origin.ConclusionThese findings indicate that a human T Cell lymphoma developed in place of this neuroblastoma PDX. Changes in PDX identity such as this one will significantly impact studies utilizing pediatric PDXs and the mechanism by which this occurred warrants further investigation.     

MAGE-1抗原肽致敏DC活化淋巴细胞对裸鼠肝癌移植瘤的免疫防护作用

目的　观察黑色素瘤 1基因 (MAGE 1)抗原肽致敏树突状细胞 (DC)所活化的淋巴细胞(CTL)对人肝癌HCC移植瘤的抑制和消退作用 ,评估临床治疗HCC的可行性和有效性。　方法BEL 74 0 2HCC细胞于 30只裸鼠背部皮下接种 ,建立裸鼠HCC移植瘤模型 ,其中 2 2只成瘤 ;用MAGE 1九肽致敏DC所活化的淋巴细胞 (1× 10 6)注入肿瘤部位皮下 (治疗组A ,n =5 ) ,其余 17只随机分成 5组 (B、C、D、E、F) ,用其他不同性质的细胞治疗 ,观察各组肿瘤生长情况并进行病理学分析和统计学处理 ,阐明特异性CTL对肿瘤的作用机制。　结果　 (1)A组HCC移植瘤均停止生长并趋于缩小 ,荷瘤裸鼠观察期内无死亡 ;而其他 5组肿瘤均快速生长 ,大部分荷瘤裸鼠 2周内死亡。MAGE 1九肽致敏DC所活化淋巴细胞能显著抑制HCC移植瘤生长 ,促使肿瘤消退 (P <0 0 1)。 (2 )A组移植肿瘤广泛坏死 ,肿瘤细胞广泛凋亡。　结论　MAGE 1九肽致敏DC有抑制HCC生长 ,促进HCC消退 ,防止肿瘤转移、复发的作用。肿瘤细胞凋亡增强是DC肿瘤免疫的可能机制。MAGE 1九肽联合DC可作为治疗HCC的新型疫苗。